Chemometric Characterization and Classification of Selected Freshwater and Marine Fishes from Turkey Based on their Fatty Acid Profiles

The fatty acid (FA) profiles of edible muscle of selected commercially important freshwater and marine fish species from Turkey were investigated. The fatty acid compositions of freshwater fish species were 23.00–29.60% saturated (SFA), 25.70–36.50% monounsaturated (MUFAs), and 26.59–31.92% polyunsaturated acids (PUFAs), whereas the fatty acid compositions of marine fish consisted of 21.08–36.90% (SFA), 18.03–51.45% MUFAs, and 20.92–53.17% PUFAs. There was a wide variation and significant (P < 0.05) differences among the fatty acid profiles of the freshwater and marine fish samples, including differences in the SFA, MUFA, PUFA, EPA, DHA, DHA/EPA, total n-3 PFAs, total n-6 PUFAs and n-3/n-6 values. In addition, the cheap marine fish species such as anchovy and European pilchard, bogue are better dietary sources of n-3 PUFAs than more expensive species such as bluefish, Atlantic mackerel, sea bream and sea bass. Through the application of two multivariate statistical methods, Principal Component and Hierarchical Analysis, fish species from Turkey waters were classified according to the geographical locations categorized in terms of fatty acid profiles. Clustering by fish species also gave rise to defined groups.     

Effect of freezing on quality of sea bass and gilthead sea bream

Freezing is an efficient method of fish preservation. The aim of the present work was to examine the impact of freezing in fatty acid composition and in the in vitro inhibitory activity of sea bass (Dicentrarchus labrax) and gilthead sea bream (Sparus aurata) fillet lipids against platelet activating factor (PAF). The Bligh and Dyer extraction method and the counter‐current distribution method were used to obtain total, polar and neutral lipids. The fatty acid analysis conducted using the internal standard method and the biological assay on washed rabbit platelets took place calculating the in vitro inhibitory activity of fish lipids against 2.5 × 10^?11 M of PAF. No statistical changes (p<0.05) occurred in fatty acid content of fresh and thawed gilthead sea bream, while fatty acid amount in thawed sea bass was significantly higher (p<0.05) compared to fresh fish. Total lipids of both thawed fish species exhibited stronger anti‐thrombotic activity compared to fresh fish. Freezing preserved fish quality and reinforced the anti‐thrombotic properties of fish oils, since even after 6 months of freezing, fish oils preserve their nutritional value in terms of protecting against cardiovascular diseases. Practical applications: Fish fillets contain high amount of unsaturated lipids that may easily undergo lipid oxidation. Freezing and frozen storage prevent such oxidative changes so fish quality is retained. Fatty acids and PAF‐antagonists in fish are of major importance since they contribute to the nutritional value of fish. The practical application of this work lies on the evaluation of the nutritional value of fish in terms of cardio protection by examining the impact of freezing on the levels of fatty acids and PAF‐antagonists in aquacultured fish fillets.     

In situ synthesis of fluorapatite-ZnO nanocomposite powder via mechanical alloying for biomedical applications

This research aimed to synthesize of fluorapatite(FA)–ZnO (FAZ) nanocomposite powder using a one-step mechanical alloying process without any post-calcification process. Furthermore, the role of various concentrations of ZnO (0, 4, 8, 11, and 15 wt.%) on the chemical and biological characteristics of FA was examined. Our research confirmed the successful synthesize of FAZ nanopowders with irregular-shaped particles and particle size of 38-52 nm. The crystallinity, crystallite size, and c-axes parameter of FA decreased with increasing ZnO content. Moreover, while all FAZ powders revealed superior bioactivity than FA, FA-8 wt.% ZnO nanopowder significantly enhanced MG63 cell proliferation. Besides, between various samples, FA-11 wt.% ZnO nanopowder revealed drastically greater antibacterial activity. It could be concluded that the in-situ synthesis of FAZ powder meaningfully improved the biological activity of FA making it a promising coating for orthopedic and dental implants.     

Preclinical Development of FA5, a Novel AMP-Activated Protein Kinase (AMPK) Activator as an Innovative Drug for the Management of Bowel Inflammation

Acadesine (ACA), a pharmacological activator of AMP-activated protein kinase (AMPK), showed a promising beneficial effect in a mouse model of colitis, indicating this drug as an alternative tool to manage IBDs. However, ACA displays some pharmacodynamic limitations precluding its therapeutical applications. Our study was aimed at evaluating the in vitro and in vivo effects of FA-5 (a novel direct AMPK activator synthesized in our laboratories) in an experimental model of colitis in rats. A set of experiments evaluated the ability of FA5 to activate AMPK and to compare the efficacy of FA5 with ACA in an experimental model of colitis. The effects of FA-5, ACA, or dexamethasone were tested in rats with 2,4-dinitrobenzenesulfonic acid (DNBS)-induced colitis to assess systemic and tissue inflammatory parameters. In in vitro experiments, FA5 induced phosphorylation, and thus the activation, of AMPK, contextually to the activation of SIRT-1. In vivo, FA5 counteracted the increase in spleen weight, improved the colon length, ameliorated macroscopic damage score, and reduced TNF and MDA tissue levels in DNBS-treated rats. Of note, FA-5 displayed an increased anti-inflammatory efficacy as compared with ACA. The novel AMPK activator FA-5 displays an improved anti-inflammatory efficacy representing a promising pharmacological tool against bowel inflammation.     

Authenticating Production Origin of Gilthead Sea Bream (<Emphasis Type="Italic">Sparus aurata)</Emphasis> by Chemical and Isotopic Fingerprinting

Recent EU legislation (EC/2065/2001) requires that fish products, of wild and farmed origin, must provide consumer information that describes geographical origin and production method. The aim of the present study was to establish methods that could reliably differentiate between wild and farmed European gilthead sea bream (Sparus aurata). The methods that were chosen were based on chemical and stable isotopic analysis of the readily accessible lipid fraction. This study examined fatty acid profiles by capillary gas chromatography and the isotopic composition of fish oil (δ¹³C, δ¹⁸O), phospholipid choline nitrogen (δ¹⁵N) and compound specific analysis of fatty acids (δ¹³C) by isotope ratio mass spectroscopy as parameters that could reliably discriminate samples of wild and farmed sea bream. The sample set comprised of 15 farmed and 15 wild gilthead sea bream (Sparus aurata), obtained from Greece and Spain, respectively. Discrimination was achieved using fatty acid compositions, with linoleic acid (18:2n-6), arachidonic acid (20:4n-6), stearic acid (18:0), vaccenic acid (18:1n-7) and docosapentaenoic acid (22:5n-3) providing the highest contributions for discrimination. Principle components analysis of the data set highlighted good discrimination between wild and farmed fish. Factor 1 and 2 accounted for >70% of the variation in the data. The variables contributing to this discrimination were: the fatty acids 14:0, 16:0, 18:0, 18:1n-9, 18:1n-7, 22:1n-11, 18:2n-6 and 22:5n-3; δ¹³C of the fatty acids 16:0, 18:0, 16:1n-7, 18:1n-9, 20:5n-3 and 22:6n-3; Bulk oil fraction δ¹³C; glycerol/choline fraction bulk δ¹³C; δ¹⁵N; % N; % lipid.     

Replacement of Marine Fish Oil with de novo Omega-3 Oils from Transgenic Camelina sativa in Feeds for Gilthead Sea Bream (Sparus aurata L.)

Omega‐3 (n‐3) long‐chain polyunsaturated fatty acids (LC‐PUFA) are essential components of the diet of all vertebrates. The major dietary source of n‐3 LC‐PUFA for humans has been fish and seafood but, paradoxically, farmed fish are also reliant on marine fisheries for fish meal and fish oil (FO), traditionally major ingredients of aquafeeds. Currently, the only sustainable alternatives to FO are vegetable oils, which are rich in C₁₈ PUFA, but devoid of the eicosapentaenoic (EPA) and docosahexaenoic acids (DHA) abundant in FO. Two new n‐3 LC‐PUFA sources obtained from genetically modified (GM) Camelina sativa containing either EPA alone (ECO) or EPA and DHA (DCO) were compared to FO and wild‐type camelina oil (WCO) in juvenile sea bream. Neither ECO nor DCO had any detrimental effects on fish performance, although final weight of ECO‐fed fish (117 g) was slightly lower than that of FO‐ and DCO‐fed fish (130 and 127 g, respectively). Inclusion of the GM‐derived oils enhanced the n‐3 LC‐PUFA content in fish tissues compared to WCO, although limited biosynthesis was observed indicating accumulation of dietary fatty acids. The expression of genes involved in several lipid metabolic processes, as well as fish health and immune response, in both liver and anterior intestine were altered in fish fed the GM‐derived oils. This showed a similar pattern to that observed in WCO‐fed fish reflecting the hybrid fatty acid profile of the new oils. Overall the data indicated that the GM‐derived oils could be suitable alternatives to dietary FO in sea bream.     

Recombinant Bovine Growth Hormone-Induced Metabolic Remodelling Enhances Growth of Gilthead Sea-Bream (Sparus aurata): Insights from Stable Isotopes Composition and Proteomics

Growth hormone and insulin-like growth factors (GH/IGF axis) regulate somatic growth in mammals and fish, although their action on metabolism is not fully understood in the latter. An intraperitoneal injection of extended-release recombinant bovine growth hormone (rbGH, Posilac^®) was used in gilthead sea bream fingerlings and juveniles to analyse the metabolic response of liver and red and white muscles by enzymatic, isotopic and proteomic analyses. GH-induced lipolysis and glycogenolysis were reflected in liver composition, and metabolic and redox enzymes reported higher lipid use and lower protein oxidation. In white and red muscle reserves, rBGH increased glycogen while reducing lipid. The isotopic analysis of muscles showed a decrease in the recycling of proteins and a greater recycling of lipids and glycogen in the rBGH groups, which favoured a protein sparing effect. The protein synthesis capacity (RNA/protein) of white muscle increased, while cytochrome-c-oxidase (COX) protein expression decreased in rBGH group. Proteomic analysis of white muscle revealed only downregulation of 8 proteins, related to carbohydrate metabolic processes. The global results corroborated that GH acted by saving dietary proteins for muscle growth mainly by promoting the use of lipids as energy in the muscles of the gilthead sea bream. There was a fuel switch from carbohydrates to lipids with compensatory changes in antioxidant pathways that overall resulted in enhanced somatic growth.     

Fatty Acid Profile of Sunshine Bass: I. Profile Change is Affected by Initial Composition and Differs Among Tissues

Fatty acid (FA) composition of fillet tissue can be tailored by transitioning fish from alternative lipid-based, low long-chain polyunsaturated fatty acid (LC-PUFA) grow-out feeds to high LC-PUFA "finishing" feeds. To address whether grow-out feed composition influences the responsiveness of fillet tissue to finishing, sunshine bass (SB, Morone chrysops x M. saxatilis) were reared to a submarketable size on grow-out feeds containing fish oil (FO) or a 50:50 blend of FO and coconut (CO), grapeseed (GO), linseed (LO), or poultry (PO) oil. For the final 8 weeks of the trial, fish were either maintained on assigned grow-out feeds or finished with the 100% FO feed. Production performance was unaffected by dietary lipid source, but fillet FA profile generally conformed to nutritional history. Regardless of grow-out regimen, finishing had a significant restorative effect on fillet FA composition; however, complete restoration of control levels of 20:5n-3, 22:6n-3, total LC-PUFA and n-3:n-6 FA ratio was achieved only among fish fed the CO-based grow-out feed. Saturated fatty acids (SFA) appear to be preferential catabolic substrates, whereas medium-chain and long-chain PUFA are selectively deposited in tissues. Provision of SFA in grow-out feeds appears to optimize selective FA metabolism and restoration of beneficial fillet FA profile during finishing.     

Dietary Source of Stearidonic Acid Promotes Higher Muscle DHA Concentrations than Linolenic Acid in Hybrid Striped Bass

Rapid expansion of aquacultural production is placing increasing demand on fish oil supplies and intensified the search for alternative lipid sources. Many of the potential alternative sources contain low concentrations of long chain n-3 fatty acids and the conversion of dietary linolenic acid to longer chain highly unsaturated fatty acids is a relatively inefficient process in some species. A 6-week study was conducted to compare tissue fatty acid (FA) concentrations in hybrid striped bass fed either 18:3n-3 (α-linolenic acid; ALA) or 18:4n-3 (stearidonic acid; SDA). Hybrid striped bass were fed either a control diet containing fish oil, or diets containing ALA or SDA at three different levels (0.5, 1 and 2% of the diet). There were no significant differences in whole animal responses between fish fed ALA or SDA. Liver and muscle concentrations of ALA and SDA were responsive to dosages fed. However, only 22:6n-3 concentrations in muscle were significantly affected by dietary source of 18 carbon precursors. Muscle 22:6n-3 concentrations were significantly higher in fish fed SDA compared to fish fed ALA. Based on these data, it appears that feeding SDA can increase long chain n-3 fatty acid concentrations in fish muscle.     

Effect of fish oil on the fatty acid composition of human milk and maternal and infant erythrocytes

To examine the effect of fish oil supplementation on the fatty acid (FA) composition of human milk and maternal and infant erythrocytes, five lactating women were supplemented with 6 g of fish oil daily for 21d. Usual maternal diets contained 1,147 mg of total n−3 FA, with 120 mg from very long-chain (>C₁₈) n−3 FA. Supplementation increased dietary levels to 3,092 mg of total n−3 FA and 2,006 mg of very long-chain n−3 FA. Milk samples were collected daily, prior to fish oil ingestion, and at 4-h intervals on days 1, 7, 14 and 21. Milk n−3 FA content increased within 8 h and reached steady state levels within one week. The n−6 fatty acid content decreased. Erythrocyte eicosapentaenoic acid content increased from 0.24% to 1.4% (P<0.01) in mothers and from 0.11% to 0.70% (P<0.05) in infants. Docosapentaenoic acid increased from 1.4% to 2.2% (P<0.05) in mothers and from 0.30% to 0.78% (P<0.01) in infants. There was no significant change in docosahexaenoic acid or n−6 fatty acid content. Maternal platelet aggregation responses were variable. No differences in milk or plasma tocopherol levels were noted. Based on a paper presented at the Symposium on Milk Lipids held at the AOCS Annual Meeting, Baltimore, MD, April 1990.     

Production of PUFA Concentrates from Poultry and Fish Processing Waste

In fish and poultry processing, viscera are generally considered as a waste product and often discarded. Chicken and hilsa fish (Hilsa ilisa) viscera were used for the production of polyunsaturated fatty acids (PUFA) linoleic (18:2n-6), eicosapentaenoic (EPA; 20:5n-3) and docosahexaenoic acid (DHA; 22:6n-3). Free fatty acids (FFA) were extracted by alkaline hydrolysis of chicken and fish viscera; yields were 5.2 and 5.9% (w/w) respectively. PUFA concentrates were obtained by a two step process—deduction of saturated fatty acids (FA) by low temperature crystallization in acetone followed by urea inclusion compound-based fractionation. Acetone treatment removed 90 and 96% of saturated FA in chicken and fish viscera respectively with FA to acetone ratio of 1:12 (w/v). Using an urea to FA ratio (w/w) of 4.0, chicken viscera produced a maximum of 84.1% of PUFA concentrates containing 82.1% of linoleic acid with a yield of 10% where as in the case of fish viscera the maximum PUFA concentrates were 91.3% containing 78.2% of EPA-DHA with the yield of 11%. Thus, the utilization of poultry and fish processing waste into the production of PUFA concentrates has been shown.     

Composition and seasonal variation of fatty acids of <Emphasis Type="Italic">Diplodus vulgaris</Emphasis> L. from the Adriatic Sea

Muscle tissue from the common two-banded sea bream Diplodus vulgaris L. originating from the Adriatic Sea, Croatia, was analyzed. The FA composition of neutral (TAG) and polar (PE, PC, PI/PS) lipid classes was determined, as well as the lipid and water contents during winter and summer periods. Both the total lipid and water contents were higher in the winter period. We identified 16 different FA. The major constituents of the total FA in both seasons were saturates: palmitic (16∶0) and stearic acids (18∶0); monoenes: oleic (18∶1n−9) and palmitoleic acids (16∶1n−7); and polyunsaturates: arachidonic acid (20∶4n−6), EPA (20∶5n−3), and DHA (22∶6n−3), but their amounts and ratios differed significantly between the two seasons and between lipid fractions. The FA composition showed a noticeable pattern of seasonality that reflected fluctuations mainly in TAG. The diminution of the monounsaturated FA content in the summer was clearly followed by an increase in PUFA content. Diplodus vulgaris is a good source of natural n−3 PUFA and would therefore be suitable for inclusion in highly unsaturated low-fat diets.     

Effects of geographic source, rearing system, and season on the nutritional quality of wild and farmed Perca fluviatilis

The effects of season, geographic source (Lake Geneva, Rhine River), and rearing system (extensive, semiextensive, and intensive systems) on the lipid content and FA composition of fillets of Perca fluviatilis were studied. Significant differences in the total lipid content were found between fish coming from the Rhine River and Lake Geneva (1.21 and 1.48%, respectively). Seasonal effects were investigated quarterly for perch sampled in the Rhine River. Intensively reared perch displayed a higher lipid content (1.48%) than the other farmed perch, i.e., 1.26% for a semiextensive system and 1.16% for an extensive system. No significant difference in lipid content was found (i) between lacustrine fish and intensively reared fish or (ii) among fish from the Rhine River and the semiextensive or extensive rearing systems. The main FA were 22∶6n−3 (DHA, 21.3–37.1% of total FA), 16∶0 (17.7–20.2%), 20∶5n−3 (EPA, 9.2–13.2%), 18∶1 (8.0–11.5%), 20∶4n−6 [arachidonic acid (ARA), 1.9–10.7%], 16∶1 (4.3–6.0%), and 18∶2n−6 (2.1–6.0%). In comparison with perch coming from the Rhine River, the lacustrine fish were characterized by higher total n−6 PUFA and a lower proportion of both total monounsaturated FA (MUFA) and total n−3 PUFA. Among rearing systems, extensively farmed fish had higher n−6 PUFA and lower n−3 PUFA contents. Wild fish showed higher ARA and 18∶2n−6 than farmed fish. They also had significantly more EPA (12.5–13.2%) than farmed perch (9.2–10.9%). For DHA no difference existed between (i) the lacustrine fish (31.9% of total FA) and the intensively reared fish (33.0%) and (ii) the Rhine (37.1%) and semiextensively reared fish (36%). Effects of size, diet composition, and environmental conditions on the total lipid contents and FA composition are discussed.     

Muscle Lipids and Fatty Acid Profiles of the Sea Catfish (Arius madagascariensis) in Madagascar Inland Waters

Muscle lipids and fatty acids (FA) of catfish Arius madagascariensis were determined in catfish caught in the Betsiboka River, Madagascar, during a 5-month sampling period. Total lipids from muscle were extracted and quantified. Fatty acids were identified by means of gas chromatography–mass spectrometry of FA methyl esters and FA pyrrolidides, leading to the identification of 42 FA. Lipid content was relatively high in our fish sample and ranged from 4.3 to 6.6% of wet muscle. Three FA dominated the FA composition: palmitic acid (C16:0, 22.9–32.6%), oleic acid (C18:1n-9, 11.3–13.4%) and stearic acid (C18:0, 10.8–12.0%). A number of polyunsaturated FA (PUFA) were present in appreciable amounts, including arachidonic acid (C20:4n-6, 4.7–7.6%), docosahexaenoic acid (C22:4n-6, 3.0–8.1%), eicosapentaenoic acid (C20:5n-3, 0.6–1.0%), n-3 docosapentaenoic acid (C22:5n-3, 1.1–1.6%), n-6 docosatetraenoic acid (C22:4n-6, 0.7–1.2%) and n-6 docosapentaenoic acid (22:5n-6, 0.9–1.8%). The sum of the n-6 PUFA and n-3 PUFA was 11.3–18.8 and 7.5–13.4%, respectively. These results indicate that A. madagascariensis, an abundant freshwater fish in Madagascar rivers, may be good source of dietary PUFA.     

POLYCYCLIC AROMATIC HYDROCARBONS IN THREE VARIETIES OF FISH FROM KUWAIT BAY

The study reports levels of parent polycyclic aromatic hydrocarbons (PAHs) in three native fish species—mullet, sea bream and tonguesole—captured from Kuwait Bay (KB) area and outside the Bay from the Auha area between October 2004 and June 2005. KB has witnessed rapid urban and industrial development since the discovery of oil and is considered to be the most productive area in the region. Whole fish analysis revealed that the lower molecular weight PAHs (pyrene, fluoranthene and phenanthrene) were present in all the three varieties of fish in considerably higher amounts. Other low molecular weight PAHs like acenaphthylene, acenaphthene and anthracene were also present but in lower concentrations. Among the higher molecular weight PAHs, the frequency of detection of benzo(a)anthracene and chrysene was high and were present in high concentrations, whereas Benzo(k)fluoranthene and benzo(a)pyrene were not frequent in detection. The contamination of Σ PAHs was higher in the winter season in mullet and sea bream but in tonguesole a reverse pattern was found. Tonguesole from KB had the highest Σ PAHs suggesting that this bottom dwelling species can be used as an indicator of sediment pollution in the area. The observed seasonality in ∑ PAHs concentration in fish coupled with species difference may be related to difference in metabolic disposition of pollutants and their habitats.     

Effect of Feeding a Low Level of Encapsulated Fish Oil to Dairy Goats on Milk Yield,Composition, and Fatty Acid Profile

Contradictory results have been found in the response of dairy goats to the supplementation of fish oil in their diet to improve the n-3 polyunsaturated fatty acids (PUFA) in milk. The responses to the inclusion of fish oil in the diet of ruminants either induced milk fat depression, increased milk fat content, and/or negatively impacted milk yield. The objective of this study was to determine whether including a low dose of encapsulated fish oil in the diet of goats can modify yield, milk composition, and the fatty acid (FA) profile of milk. Ten Alpine goats were divided into two homogeneous subgroups and assigned to either the control or experimental diet. The control animals received the basal diet without supplementation of fish oil, whereas the experimental group was given the same basal diet supplemented with encapsulated fish oil (1.14 g /kg of concentrate) for 56 days. Milk samples were analyzed for chemical composition and FA profile. The inclusion of encapsulated fish oil in the goat diet did not affect the yield and composition of goat milk. The effect of diet was not significant on the FA profile of goat milk, except that 20:0 was lower (P < 0.05) in the milk of goats that received fish oil. The low dose of encapsulated fish oil supplement used in this study did not impact (P > 0.05) the PUFA content of goat milk or milk composition and yield; however, the atherogenicity index (AI), which is beneficial to heart health, was lower (P < 0.05) in the milk of goats that received fish oil as a supplement in their diet compared to the control.     

Fatty Acid Composition of Tropical Fish Depends on Reservoir Trophic Status and Fish Feeding Habit

Eutrophication results in a deficiency of n‐3 LC‐PUFA (long‐chain polyunsaturated fatty acids) in aquatic food chains, affecting fish nutrition and physiology. The trophic transfer of FA (fatty acids) to fish species of different feeding habits was investigated in two reservoirs in southeast Brazil—the mesotrophic Ponte Nova Reservoir (PN) and the hypereutrophic Billings Reservoir (Bil). Total FA profile of stomach contents and adipose tissue, triacylglycerols (TAG), and phospholipids (PL) from liver and muscle of the omnivorous Astyanax fasciatus and the carnivorous Hoplias malabaricus were analyzed by gas chromatography. A prevalence of n‐6PUFA, as 18:2n‐6 (linoleic acid) and 20:4n‐6 (arachidonic acid, ARA) was observed in the stomach contents and in the tissues of A. fasciatus from the PN reservoir. In contrast, n‐3 LC‐PUFA, as 20:5n‐3 (eicosapentaenoic acid, EPA) was accumulated in fish tissues from Bil, resulting in higher n3/n6 and EPA/ARA ratios, compared to fish from PN. This differential FA accumulation was also observed for H. malabaricus, but differences were slightly minor, and no changes were observed in the EPA/ARA ratios between fish from both reservoirs. Regardless reservoir, FA profiles of TAG resembled that of their diet, whereas FA profiles of PL were more conservative and mainly comprised by LC‐PUFA. We conclude that reservoir trophic status affected the FA composition of food resources available to these fish species, resulting in differential allocation of n‐3 and n‐6 FA. As expected, FA profile of the investigated fish species also reflected their feeding habit and physiological demands.